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| United States Patent |
5,705,502
|
|
Zimmermann
|
January 6, 1998
|
Pharmacologically active pyrimidineamine derivatives and processes for
the preparation thereof
Abstract
Described are N-phenyl-2-pyrimidineamine derivatives of formula I
##STR1##
wherein R.sub.1 is a substituted cyclic radical, the cyclic radical being
bonded at a ring carbon atom in each case and being selected from phenyl,
pyridyl, pyrazinyl, thiazolyl, pyrimidinyl, pyridazinyl and imidazolyl,
and the substituents of the above-mentioned cyclic radical being selected
from one or more of the groups halogen, cyano, carbamoyl,
--C(.dbd.O)--OR.sub.3, --C(.dbd.O)--R.sub.4, --SO.sub.2
--N(R.sub.5)--R.sub.6, --N(R.sub.7)--R.sub.8, --OR.sub.9 and
fluorine-substituted lower alkyl, wherein
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are each
independently of the others hydrogen or lower alkyl that is unsubstituted
or substituted by mono- or di-lower alkylamino; and
R.sub.2 is selected from halogen, cyano, carbamoyl, --C(.dbd.O)--OR.sub.10,
--C(.dbd.O)--R.sub.11, --SO.sub.2 --N(R.sub.12)--R.sub.13,
--N(R.sub.14)--R.sub.15, --OR.sub.16 and fluorine-substituted lower alkyl,
wherein
R.sub.10, R.sub.11, R.sub.12, R.sub.13, R.sub.14, R.sub.15 and R.sub.16 are
each independently of the others hydrogen or lower alkyl that is
unsubstituted or substituted by mono- or di-lower alkylamino. Those
compounds can be used, for example, in the treatment of tumour diseases.
| Inventors:
|
Zimmermann; Jurg (Wallbach, CH)
|
| Assignee:
|
Novartis Corporation (Summit, NJ)
|
| Appl. No.:
|
446742 |
| Filed:
|
May 31, 1995 |
| PCT Filed:
|
September 21, 1994
|
| PCT NO:
|
PCT/EP94/03148
|
| 371 Date:
|
May 31, 1995
|
| 102(e) Date:
|
May 31, 1995
|
| PCT PUB.NO.:
|
WO95/09851 |
| PCT PUB. Date:
|
April 13, 1995 |
Foreign Application Priority Data
| Oct 01, 1993[CH] | 2968/93 |
| Jul 18, 1994[CH] | 2280/94 |
| Current U.S. Class: |
514/275; 514/252.02; 514/255.05; 544/238; 544/295; 544/296; 544/331 |
| Intern'l Class: |
C07D 239/42; A61K 031/505 |
| Field of Search: |
514/252,275
544/295,331
|
References Cited
U.S. Patent Documents
| 5612340 | Mar., 1997 | Zimmermann | 514/252.
|
Primary Examiner: Ford; John M.
Attorney, Agent or Firm: Ferraro; Gregory D.
Claims
What is claimed is:
1. An N-phenyl-2-pyrimidineamine derivative of formula I
##STR9##
wherein R.sub.1 is a substituted cyclic radical, the cyclic radical being
bonded at a ring carbon atom in each case and being selected from pyridyl,
pyrazinyl, thiazolyl, pyrimidinyl, pyridazinyl and imidazolyl, and the
substituents of the above-mentioned cyclic radical being selected from one
or more of the groups halogen, cyano, carbamoyl, --C(.dbd.O)--OR.sub.3,
--C(.dbd.O)--R.sub.4, --SO.sub.2 --N(R.sub.5)--R.sub.6,
--N(R.sub.7)--R.sub.8, --OR.sub.9 and fluorine-substituted lower alkyl,
wherein
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are each
independently of the others hydrogen or lower alkyl that is unsubstituted
or substituted by mono- or di-lower alkylamino; and
R.sub.2 is selected from halogen, cyano, carbamoyl, --C(.dbd.O)--OR.sub.10,
--C(.dbd.O)--R.sub.11, --SO.sub.2 --N(R.sub.12)--R.sub.13,
--N(R.sub.14)--R.sub.15, --OR.sub.16 and fluorine-substituted lower alkyl,
wherein
R.sub.10, R.sub.11, R.sub.12, R.sub.13, R.sub.14, R.sub.15 and R.sub.16 are
each independently of the others hydrogen or lower alkyl that is
unsubstituted or substituted by mono- or di-lower alkylamino, or a salt of
such a compound having at least one salt-forming group.
2. A compound according to claim 1 of formula I, wherein
R.sub.1 is substituted pyridyl bonded to a ring carbon atom, the
substituents of the above-mentioned pyridyl radical being selected from
halogen, cyano, carbamoyl, --C(.dbd.O)--OR.sub.3, --N(R.sub.7)--R.sub.8
and --OR.sub.9, wherein
R.sub.3, R.sub.7, R.sub.8 and R.sub.9 are each independently of the others
hydrogen or lower alkyl; and
R.sub.2 is selected from halogen, --C(.dbd.O)--OR.sub.10, wherein
R.sub.10 is hydrogen or lower alkyl,
and from fluorine-substituted lower alkyl,
or a salt of such a compound having at least one salt-forming group.
3. A compound according to claim 1 of formula I, wherein
R.sub.1 is a pyridyl radical substituted by halogen, cyano, carboxy,
carbamoyl, hydroxy or by N-lower alkyl-amino, and
R.sub.2 is halogen or fluorine-substituted lower alkyl,
or a salt thereof.
4. A compound according to claim 1 of formula I, wherein
R.sub.1 is a 4-pyridyl radical substituted in the 2-position with respect
to the pyridine nitrogen by chlorine, cyano, carboxy, carbamoyl, hydroxy
or by N-propyl-amino and
R.sub.2 is chlorine or trifluoromethyl,
or a salt thereof.
5. A compound according to claim 1 of formula I, wherein
R.sub.1 is a 4-pyridyl radical substituted in the 2-position with respect
to the pyridine nitrogen by chlorine, cyano, carboxy, carbamoyl, hydroxy,
amino, N-propyl-amino, N,N-dimethylamino or by N-butyl-amino, and
R.sub.2 is chlorine, trifluoromethyl, carboxy or lower alkoxycarbonyl,
or a salt thereof.
6. A compound according to claim 1 of formula I or a pharmaceutically
acceptable salt of such a compound having at least one salt-forming group,
selected from
N-(3-chloro-phenyl)-4-(2-chloro4-pyridyl)-2-pyrimidineamine,
N-(3-trifluoromethyl-phenyl)-4-(2-chloro-4-pyridyl)-2-pyrimidineamine,
N-(3-trifluoromethyl-phenyl)-4-(2-cyano-4-pyridyl)-2-pyrimidineamine,
N-(3-trifluoromethyl-phenyl)-4-(2-carboxy-4-pyridyl)-2-pyrimidineamine,
N-(3-chloro-phenyl)-4-(2-cyano-4-pyridyl)-2-pyrimidineamine,
N-(3-chloro-phenyl)-4-(2-carboxy-4-pyridyl)-2-pyrimidineamine,
N-(3-chloro-phenyl)-4-(2-carbamoyl-4-pyridyl)-2-pyrimidineamine,
N-(3-trifluoromethyl-phenyl)-4-(2-carbamoyl-4-pyridyl)-2-pyrimidineamine,
N-(3-chloro-phenyl)-4-(2-n-propylamino-4-pyridyl)-2-pyrimidineamine,
N-(3-chloro-phenyl)-4-(2-amino-4-pyridyl)-2-pyrimidineamine,
N-(3-chloro-phenyl)-4-(2-hydroxy-4-pyridyl)-2-pyrimidineamine,
N-(3-chloro-phenyl)-4-(2-dimethylamino-4-pyridyl)-2-pyrimidineamine,
N-›3-ethoxycarbonyl-phenyl!-4-(2-chloro-4-pyridyl)-2-pyrimidineamine,
N-›3-isopropoxycarbonyl-phenyl!-4-(2-chloro-4-pyridyl)-2-pyrimidineamine,
N-›3-carboxy-phenyl!-4-(2-chloro-4-pyridyl)-2-pyrimidineamine and
N-›3-chloro-phenyl!-4-›2-(n-1-butylamino)-4-pyridyl!-2-pyrimidineamine,
and from the pharmaceutically acceptable salts of such compounds having at
least one salt-forming group.
7. A compound of formula I according to claim 1, or a pharmaceutically
acceptable salt thereof, said compound being
N-(3-chloro-phenyl)-4-(2-amino-4-pyridyl)-2-pyrimidineamine.
8. A pharmaceutical composition comprising a compound of formula I
according to claim 1 or a pharmaceutically acceptable salt of such a
compound having at least one salt-forming group together with a
pharmaceutical carrier.
9. A process for the preparation of an N-phenyl-2-pyrimidineamine
derivative of formula I
##STR10##
wherein R.sub.1 is a substituted cyclic radical, the cyclic radical being
bonded at a ring carbon atom in each case and being selected from pyridyl,
pyrazinyl, thiazolyl, pyrimidinyl, pyridazinyl and imidazolyl, and the
substituents of the above-mentioned cyclic radical being selected from one
or more of the groups halogen, cyano, carbamoyl, --C(.dbd.O)--OR.sub.3,
--C(.dbd.O)--R.sub.4, --SO.sub.2 --N(R.sub.5)--R.sub.6,
--N(R.sub.7)--R.sub.8, --OR.sub.9 and fluorine-substituted lower alkyl,
wherein
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are each
independently of the others hydrogen or lower alkyl that is unsubstituted
or substituted by mono- or di-lower alkylamino; and
R.sub.2 is selected from halogen, cyano, carbamoyl, --C(.dbd.O)--OR.sub.10,
--C(.dbd.O)--R.sub.11, --SO.sub.2 --N(R.sub.12)--R.sub.13,
--N(R.sub.14)--R.sub.15, --OR.sub.16 and fluorine-substituted lower alkyl,
wherein
R.sub.10, R.sub.11, R.sub.12, R.sub.13, R.sub.14, R.sub.15 and R.sub.16 are
each independently of the others hydrogen or lower alkyl that is
unsubstituted or substituted by mono- or di-lower alkylamino, or of a salt
of such a compound having at least one salt-forming group, wherein a
compound of formula II
R.sub.1 --C(.dbd.O)--CH.dbd.CH--N(R.sub.17)--R.sub.18 (II),
wherein R.sub.17 and R.sub.18 are each independently of the other lower
alkyl and R.sub.1 is as defined above, functional groups present in a
compound of formula II, with the exception of the groups participating in
the reaction, being, if necessary, in protected form, or a salt of such a
compound is reacted with a compound of formula III
##STR11##
wherein R.sub.2 is as defined above, functional groups present in a
compound of formula III, with the exception of the guanidino group
participating in the reaction, being, if necessary, in protected form, or
with a salt of such a compound, and any protecting groups present are
removed
and, if desired, a compound of formula I obtainable in accordance with the
above process is converted into its salt, or an obtainable salt of a
compound of formula I is converted into the free compound.
Description
This application is filed under 35 U.S.C. .sctn. 371 as a national phase
application of PCT/EP94/03148, filed Sep. 21, 1994 published as
WO95/09851, Apr. 13, 1995.
The invention relates to N-phenyl-2-pyrimidineamine derivatives, to
processes for the preparation thereof, to medicaments comprising those
compounds, and to the use thereof in the preparation of pharmaceutical
compositions for the therapeutic treatment of warm-blooded animals.
The invention relates to N-phenyl-2-pyrimidineamine derivatives of formula
I
##STR2##
wherein R.sub.1 is a substituted cyclic radical, the cyclic radical being
bonded to a ring carbon atom in each case and being selected from phenyl,
pyridyl, pyrazinyl, thiazolyl, pyrimidinyl, pyridazinyl and imidazolyl,
and the substituents of the above-mentioned cyclic radical being selected
from one or more of the groups halogen, cyano, carbamoyl,
--C(.dbd.O)--OR.sub.3, --C(.dbd.O)--R.sub.4, --SO.sub.2
--N(R.sub.5)--R.sub.6, --N(R.sub.7)--R.sub.8, --OR.sub.9 and
fluorine-substituted lower alkyl, wherein
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are each
independently of the others hydrogen or lower alkyl that is unsubstituted
or substituted by mono- or di-lower alkylamino; and
R.sub.2 is selected from halogen, cyano, carbamoyl, --C(.dbd.O)--OR.sub.10,
--C(.dbd.O)--R.sub.11, --SO.sub.2 --N(R.sub.12)--R.sub.13,
--N(R.sub.14)--R.sub.15, --OR.sub.16 and fluorine-substituted lower alkyl,
wherein
R.sub.10, R.sub.11, R.sub.12, R.sub.13, R.sub.14, R.sub.15 and R.sub.16 are
each independently of the others hydrogen or lower alkyl that is
unsubstituted or substituted by mono- or di-lower alkylamino, and to salts
of such compounds having at least one salt-forming group.
A substituted cyclic radical R.sub.1, such as, for example, a substituted
phenyl radical R.sub.1, can have several substituents, but especially not
more than 3 and, especially in the case of relatively large substituents,
preferably only one substituent, which substituents are principally in the
para- (or 4-position) and/or preferably meta-position (or 3-position) with
respect to the bonding site of the cyclic radical R.sub.l. The
above-mentioned substituted cyclic radicals R.sub.1 other than phenyl
generally have up to two and preferably only one substituent, which is/are
especially in the para-position and/or preferably meta-position with
respect to the bonding site of the cyclic radical R.sub.1.
Pyridyl bonded to a ring carbon atom is 2- or preferably 4- or 3-pyridyl,
especially 4-pyridyl. In a mono-substituted pyridyl radical R.sub.1, the
substituent is preferably in the ortho-position with respect to the
pyridine nitrogen.
Halogen in a radical R.sub.1 is preferably chlorine or fluorine.
Halogen-substituted phenyl R.sub.1 is preferably 2-, 3- or 4-chloro-phenyl,
2,4-, 3,4- or 2,5-dichloro-phenyl or 2,3,4-trichloro-phenyl.
Fluorine-substituted lower alkyl R.sub.2 is lower alkyl that carries at
least one, but preferably several, fluorine substituents, especially
1,1,2,2-tetrafluoro-ethyl or more especially tri-fluoromethyl.
Mono- or di-lower alkylamino is, for example, methylamino or dimethylamino.
Within the scope of this text, the term "lower" denotes radicals having up
to and including 7, preferably up to and including 4, carbon atoms.
Unless otherwise indicated in the context concerned, lower alkyl is
preferably methyl or ethyl.
R.sub.3 and R.sub.7 are preferably hydrogen. R.sub.8 is preferably lower
alkyl, such as especially n-propyl. R.sub.9 is preferably hydrogen or
methyl.
Salt-forming groups in a compound of formula I are groups or radicals
having basic or acidic properties. Compounds having at least one basic
group or at least one basic radical, for example a mono-lower alkylamino
group, a pyrazinyl radical or a pyridyl radical, can form acid addition
salts, for example with inorganic acids, such as hydrochloric acid,
sulfuric acid or a phosphoric acid, or with suitable organic carboxylic or
sulfonic acids, for example aliphatic mono- or di-carboxylic acids, such
as trifluoroacetic acid, acetic acid, propionic acid, glycolic acid,
succinic acid, maleic acid, fumaric acid, hydroxymaleic acid, malic acid,
tartaric acid, citric acid, oxalic acid or amino acids, such as arginine
or lysine, aromatic carboxylic acids, such as benzoic acid,
2-phenoxy-benzoic acid, 2-acetoxy-benzoic acid, salicylic acid,
4-aminosalicylic acid, aromatic-aliphatic carboxylic acids, such as
mandelic acid or cinnamic acid, heteroaromatic carboxylic acids, such as
nicotinic acid or isonicotinic acid, aliphatic sulfonic acids, such as
methane-, ethane- or 2-hydroxy-ethane-sulfonic acid, or aromatic sulfonic
acids, for example benzene-, p-toluene- or naphthalene-2-sulfonic acid. If
several basic groups are present, mono- or poly-acid addition salts can be
formed.
Compounds of formula I having acidic groups, for example a free carboxy
group in the radical R.sub.1, can form metal or ammonium salts, such as
alkali metal or alkaline earth metal salts, for example sodium, potassium,
magnesium or calcium salts, or ammonium salts with ammonia or suitable
organic amines, such as tertiary monoamines, for example tri-ethylamine or
tri(2-hydroxyethyl)amine, or heterocyclic bases, for example
N-ethyl-piperidine or N,N'-dimethyl-piperazine.
Compounds of formula I that possess both acidic and basic groups can form
internal salts.
For the purpose of isolation or purification and also in the case of the
compounds used further as intermediates, it is also possible to use
pharmaceutically unacceptable salts. Only the pharmaceutically acceptable
non-toxic salts are used therapeutically, however, and those are therefore
preferred.
In view of the close relationship between the novel compounds in free form
and in the form of their salts, including also salts that can be used as
intermediates, for example in the purification of the novel compounds or
in order to identify those compounds, hereinbefore and hereinafter any
reference to the free compounds is to be understood as including also the
corresponding salts, where appropriate and expedient.
The compounds of formula I exhibit valuable pharmacological properties: for
example, they inhibit the enzyme protein kinase C with a high degree of
selectivity. Phospholipid- and calcium-dependent protein kinase C occurs
in cells in a number of forms and participates in various fundamental
processes, such as signal transmission, proliferation and differentiation,
and also the release of hormones and neurotransmitters. The activation of
that enzyme is effected either by receptor-mediated hydrolysis of
phospholipids of the cell membrane or by direct interaction with certain
turnout-promoting active substances. The sensitivity of the cell to
receptor-mediated signal transmission can be substantially influenced by
modifying the activity of protein kinase C (as a signal transmitter).
Compounds that are capable of influencing the activity of protein kinase C
can be used as tumour-inhibiting, an antiinflammatory, immunomodulating
and antibacterial active ingredients and may even be of value as agents
against atherosclerosis and disorders of the cardiovascular system and
central nervous system.
Formerly, porcine brain protein kinase C purified in accordance with the
procedure described by T. Uchida and C. R. Filburn in J. Biol. Chem. 259,
12311-4 (1984) was used to determine the inhibitory action on protein
kinase C, and the inhibitory action on protein kinase C was determined in
accordance with the procedure of D. Fabbro et al., Arch. Biochem. Biophys.
239, 102-111 (1985).
The porcine brain protein kinase C formerly used is a mixture of various
sub-types (isotypes) of protein kinase C. If pure recombinant isotypes are
used instead of porcine brain protein kinase C in the above test it is
found that the compounds of formula I inhibit the "conventional" isotype
.alpha. preferentially whereas the other "conventional" isotypes .beta.-1,
.beta.-2 and .gamma. and especially the "non-conventional" isotypes
.delta., .epsilon. and .eta. and the "atypical" isoform .zeta. are
generally inhibited to a distinctly lesser extent and in some cases hardly
at all.
Recombinant PKC isotypes are cloned, expressed and purified in the
following manner:
The production of various proteins with the aid of baculoviruses, and their
cloning and isolation from Sf9 insect cells are carried out as described
by M. D. Summers and G. E. Smith, "A manual method for baculovirus vectors
and insect cell culture procedure", Texas Agricul. Exptl. Station Bull
(1987), 1555. The construction and isolation of recombinant viruses for
the expression of PKC-.alpha.(bovine), PKC-.beta.1(human),
PKC-.beta.2(human) and PKC-.gamma.(human/bovine hybrid) in Sf9 cells are
effected in the manner described by Stabel et al ›S. Stabel, M. Liyanage
and D. Frith, "Expression of protein kinase C isozymes in insect cells and
isolation of recombinant proteins", Meth. Neurosc. (1993)!. The production
of the PKC isotypes in Sf9 cells is carried out in the manner indicated by
Stabel et al. (see above), and the purification of the enzymes is effected
in accordance with the method described in the publication by McGlynn et
al. ›E. McGlynn, J. Liebetanz, S. Reutener, J. Wood, N. B. Lydon, H.
Hofstetter, M. Vanek, T. Meyer and D. Fabbro, "Expression and partial
characterization of rat protein kinase C-.delta. and protein kinase
C-.zeta. in insect cells using recombinant baculovirus", J. Cell Biochem.
49, 239-250 (1992)!. For the generation of recombinant PKC-.delta.(rat),
PKC-.epsilon.(rat), PKC-.zeta.(rat) and PKC-.eta.(mouse), and their
expression and purification, the procedure described by Liyanage et al
›"Protein kinase C group B members PKC-.delta., -.epsilon., -.zeta. and
PKC-.lambda.: Comparison of properties of recombinant proteins in vitro
and in vivo", Biochem. J. 283, 781-787 (1992)! and McGlynn et al,
respectively, (see above) is followed, with the additional feature that
the transfer vector pAc360 is used for the expression of PKC-.eta.›V.
Luckow and M. D. Summers, "Trends in the development of baculovirus
expression", Biotechnology 6, 47-55 (1988)!.
The measurement of the activity of the recombinant PKC isotypes obtained by
the above method is carried out in the absence of lipid and calcium
(co-factors). Protamine sulfate phosphorylated in the absence of
co-factors is used as the substrate. The activity of the enzymes reflects
the transfer of .sup.32 P from .gamma.›.sup.32 P!-ATP to protamine
sulfate. Protamine sulfate is a mixture of polypeptides each comprising
four C-terminal arginine residues. Phosphate incorporation is measured
under the following conditions: 100 .mu.l of the reaction mixture comprise
in final concentrations 20 mM TRIS-HCl pH 7.4, 10 mM Mg›NO.sub.3 !.sub.2,
0.5 mg/ml of protamine sulfate, 10 .mu.M ATP (0.1 .mu.Ci .gamma.-›.sup.32
P!-ATP; 10 Ci/mol; Amersham, Little Chalfont, United Kingdom), various
concentrations of the inhibitory compounds and 0.5-2.5 U (units: a unit is
the mount of enzyme that, in one minute and per milligram of protein,
transfers one nanomole of .sup.32 P from the above-mentioned
.gamma.-›.sup.32 P!-ATP to histone H1 ›Sigma, type V-S!) of the enzymes.
The reaction is started by the addition of the enzymes and transfer at
32.degree. C. The reaction time is 20 minutes. The reaction is then
stopped by dripping aliquots of 50 .mu.l onto P81 chromatography paper
(Whatman, Maidstone, United Kingdom). After removing unbound
.gamma.›.sup.32 P!-ATP and nucleotide fragments by washing operations as
described by J. J. Witt and R. Roskoski, "Rapid protein kinase assay using
phospho-cellulose-paper absorption", Anal. Biochem. 66, 253-258 (1975),
the substrate phosphorylation is determined by scintillation measurement.
In that test, the compounds of formula I inhibit the .alpha.-isotype of
protein kinase C (PKC) at an IC.sub.50 of as low as approximately from 1
to 75 .mu.mol/liter, generally approximately from 1 to 10 .mu.mol/liter.
In contrast, the other isotypes of PKC are generally inhibited only at
distinctly higher concentrations (i.e. at concentrations up to more than
300 times higher).
As may be expected purely on the basis of the above-described inhibitory
action on protein kinase C, the compounds of formula I exhibit
antiproliferative properties which can be demonstrated directly in another
test described in the following in which the inhibitory action of the
compounds of formula I on the growth of human T24 bladder carcinoma cells
is determined. Those cells are incubated in Eagle's minimal essential
medium, to which 5% (v/v) foetal calf serum has been added, in a
humidified incubator at 37.degree. C. and with 5% by volume of CO.sub.2 in
the air. The carcinoma cells (1000-1500) are sown in 96-well microtitre
plates and incubated overnight under the above-mentioned conditions. The
test compound is added in serial dilutions on day 1. The plates are
incubated for 5 days under the above-mentioned conditions. During that
period the control cultures undergo at least four cell divisions. After
incubation, the cells are fixed with 3.3% (w/v) aqueous glutaraldehyde
solution, washed with water and stained with 0.05% (weight/volume) aqueous
methylene blue solution. After washing, the dye is eluted with 3% (w/v)
aqueous hydrochloric acid. The optical density (OD) per well, which is
directly proportional to the number of cells, is then measured at 665 nm
using a photometer (Titertek multiskan). The IC.sub.50 values are
calculated with a computer system using the formula
##EQU1##
The IC.sub.50 values are defined as being the concentration of active
ingredient at which the number of cells per well at the end of the
incubation period is only 50% of the number of cells in the control
cultures. In the case of the compounds of formula I, the IC.sub.50 values
so ascertained are generally approximately from 1 to 20 .mu.mol/liter.
The anti-tumour activity of the compounds of formula I can also be
demonstrated in vivo:
Female Balb/c hairless mice with s.c. transplanted human bladder tumours
T24 are used to determine the anti-tumour activity. On day 0, with the
animals under peroral forene narcosis, approximately 25 mg of a solid
tumour are placed under the skin on the animals' left flank and the small
incised wound is closed by means of suture clips. On day 6 after the
transplantation, the mice are divided at random into groups of 6 animals
and treatment commences. The treatment is carried out for 15 days with
peroral or intraperitoneal administration once daily of a compound of
formula I in dimethyl sulfoxide/Tween 80/-sodium chloride solution in the
various doses. The tumours are measured twice a week with a slide gauge
and the volume of the tumours is calculated. In that test, the peroral or
intraperitoneal administration of a compound of formula I brings about a
marked reduction in the avenge tumour volume in comparison with the
untreated control animals.
On the basis of the properties described, the compounds of formula I can be
used especially as tumour-inhibiting active ingredients, for example in
the treatment of tumours of the bladder and the skin. When the compounds
of formula I are used in the treatment of cancer in combination with other
chemotherapeutic drugs, they prevent the development of resistance
(multidrug resistance) or eliminate an already existing resistance to the
other chemotherapeutic drugs. They are also suitable for the other uses
mentioned above for protein kinase C modulators and can be used especially
in the treatment of disorders responsive to inhibition of protein kinase
C.
Some of the compounds of formula I also inhibit the tyrosine kinase
activity of the receptor for the epidermal growth factor (EGF). That
receptor-specific enzyme activity plays a key role in signal transmission
in a large number of mammalian cells, including human cells, especially
epithelial cells, cells of the immune system and cells of the central and
peripheral nervous system. In the case of various types of cell, the
EGF-induced activation of the receptor-associated tyrosine protein kinase
(EGF-R-TPK) is a prerequisite for cell division and accordingly for the
proliferation of a cell population. The addition of EGF-receptor-specific
tyrosine kinase inhibitors thus inhibits the replication of those cells.
Inhibition of EGF-receptor-specific tyrosine protein kinase (EGF-R-TPK) can
be demonstrated, for example, using the method of E. McGlynn et al, Europ.
J. Biochem. 207, 265-275 (1992). The compounds according to the invention
inhibit the enzyme activity by 50% (IC50) for example at a concentration
of from 0.1 to 10 .mu.M.
The compounds of formula I that inhibit the tyrosine kinase activity of the
receptor for the epidermal growth factor (EGF) can accordingly be used,
for example, in the treatment of benign or malignant tumours. They are
able to bring about tumour regression and to prevent metastatic spread and
the growth of micrometastases. They can be used especially in the case of
epidermal hyperproliferation (psoriasis), in the treatment of neoplasia of
epithelial character, for example mastocarcinoma, and in the case of
leukaemia. The compounds can also be used in the treatment of disorders of
the immune system and inflammation if protein kinases are involved.
Furthermore, those compounds of formula I can be used in the treatment of
disorders of the central or peripheral nervous system if signal
transmission by protein kinases is involved.
The compounds of formula I and salts of such compounds having at least one
salt-forming group also inhibit the enzyme p34.sup.cdc2 /cycline
B.sup.cdc13 kinase. That kinase controls, in addition to other
cdc2-related kinases, specific phases of cell division, especially the
transition from the G.sub.1 -phase to the S-phase and more especially the
transition from the G.sub.2 -phase to the M-phase.
In chronological order, the cycle of a eukaryotic cell consists of the
interphase and the M-phase. The interphase is accompanied by an increase
in the size of the cell. In chronological order, the interphase consists
for its part of the G.sub.1 -phase, the S-phase and the G.sub.2 -phase. In
the G.sub.1 -phase (G=gap) biosynthetic processes take place in the cell.
In the S-phase (synthesis phase) the DNA doubles. The cell then enters the
G.sub.2 -phase which ends with the commencement of mitosis.
In chronological order, the M-phase for its part consists of the division
of the cell nucleus (mitosis) and the division of the cytoplasm
(cytokinesis).
The above-mentioned inhibition of the enzyme p34.sup.cdc2 /cycline
B.sup.cdc13 kinase can be demonstrated by the following test;
10 .mu.M 1-methyl-adenine are used to induce starfish oocytes to enter the
M-phase. The oocytes are then frozen in liquid nitrogen and stored at
-80.degree. C. If necessary, the oocytes are homogenised and centrifuged,
as described in D. Arion et al, Cell 55, 371-378 (1988) and V. Rialet and
L. Meijer, Anticancer Res. 11, 1581-1590 (1991). In order to purify the
p34.sup.cdc2 /cycline B.sup.cdc13 kinase, the supernatant of the oocytes
is added to p9.sup.CKShs -Sepharose grains prepared from recombinant human
protein p9.sup.CKShs, as described in L. Azzi et al., Eur. J. Biochem.
203, 353-360 (1992). After 30 minutes at 4.degree. C. while being turned
constantly, the grains are washed thoroughly and the active p34.sup.cdc2
/cycline B.sup.cdc13 kinase is eluted with free protein p9.sup.CKShs (3
mg/ml). The eluted kinase is tested using historic H1 as substrate, as
described in L. Meijer et al., EMBO J. 8, 2275-2282 (1989) and EMBO J. 10,
1545-1554 (1991). In that test, the compounds of formula I and salts of
such compounds having at least one salt-forming group exhibit an
inhibiting concentration IC.sub.50 ›.mu.mol/liter! of approximately from
0.01 to 2.
That finding would also lead to the expectation that the compounds of
formula I and salts of such compounds having at least one salt-forming
group can be used in the treatment of hyperproliferative disorders, such
as tumours and psoriasis.
The compounds of formula I also inhibit the production of HIV viruses, as
shown by the test below, and can accordingly be used as agents against the
immune deficiency disease AIDS. The initial symptoms observed after HIV
infection in humans is followed by a clinical latency period which can
last several years. After that period, the stage known as AIDS occurs and
usually progresses to death. The latency period is attributed to several
factors: immune response, occlusion of the viruses in lymph nodes or other
tissue and entry into a stage of molecular and viral latency in which the
infected cells do not complete the viral cell cycle, which is why
infectious viruses cannot be produced and the infection cannot spread.
That stage of molecular latency has been investigated using cell models,
such as the ACH-2 cell line ›K. Clouse et al., J. Immunol. 142, 431
(1989)! and the U1 cell line ›T. Folks et al., J. Immunol. 140, 117
(1988)!. Those cells are infected with HIV-1 viruses, but have only a low
content of infectious viruses. If, however, those cells are stimulated
with physiologically relevant factors that are known to be increased in
AIDS patients, such as tumour necrosis factor, interleukin-6 etc., or with
chemical inductors, such as phorbol diesters for example
13-O-acetyl-12-O-n-tetradecanoyl-phorbol, a massive production of virus
follows. The ACH-2 and U1 cells are representatives of two different cell
families that are targets for HIV infection, namely lymphocytes and
macrophages.
Hitherto, effective prevention of the progression of HIV infection to the
outbreak of AIDS has not been possible. Many attempts have been made to
prevent virus replication after the outbreak of AIDS, that is to say, in a
stage in which viruses are produced on a massive scale. In contrast, the
compounds of formula I interfere with cell processes that lead to the
activation of Intently infected HIV cells without impairing normal cell
processes, such as cell division.
If the above-mentioned U 1 or ACH-2 cells are used as a model for vital
latency, it can be demonstrated that HIV virus production induced by
13-O-acetyl-12-O-n-tetradecanoyl-phorbol or tumour necrosis factor-alpha
are effectively inhibited by the compounds of formula I at a concentration
of approximately from 0.001 to 1 .mu.mol/liter, for example at 0.03
.mu.mol/liter.
Preferred are compounds of formula I wherein
R.sub.1 is substituted pyridyl bonded to a ring carbon atom, the
substituents of the above-mentioned pyridyl radical being selected from
halogen, cyano, carbamoyl, --C(.dbd.O)--OR.sub.3, --N(R.sub.7)--R.sub.8
and --OR.sub.9, wherein
R.sub.3, R.sub.7, R.sub.8 and R.sub.9 are each independently of the others
hydrogen or lower alkyl; and
R.sub.2 is selected from halogen, --C(.dbd.O)--OR.sub.10, wherein
R.sub.10 is hydrogen or lower alkyl,
and from fluorine-substituted lower alkyl,
and salts of such compounds having at least one salt-forming group.
Preferred are especially compounds of formula I wherein
R.sub.1 is a pyridyl radical substituted by halogen, cyano, carboxy,
carbamoyl, hydroxy or by N-lower alkyl-amino, and
R.sub.2 is halogen or fluorine-substituted lower alkyl,
and the salts thereof.
Especially preferred are compounds of formula I wherein
R.sub.1 is a 4-pyridyl radical substituted in the 2-position by chlorine,
cyano, carboxy, carbamoyl, hydroxy or by N-propyl-amino and
R2 is chlorine or trifluoromethyl,
and the salts thereof.
Very preferred are also compounds of formula I wherein
R.sub.1 is a 4-pyridyl radical substituted in the 2-position with respect
to the pyridine nitrogen by chlorine, cyano, carboxy, carbamoyl, hydroxy,
amino, N-propyl-amino, N,N-dimethylamino or by N-butyl-amino, and
R.sub.2 is chlorine, trifluoromethyl, carboxy or lower alkoxycarbonyl,
and the salts thereof.
More especially preferred are the compounds of formula I described in the
Examples.
The compounds of formula I and the salts of such compounds having at least
one salt-forming group are prepared in accordance with processes known per
se. The process according to the invention is effected as follows:
a) a compound of formula II
R.sub.1 --C(.dbd.O)--CH.dbd.CH--N(R.sub.17)--R.sub.18 (II),
wherein R.sub.17 and R.sub.18 are each independently of the other lower
alkyl and R.sub.1 is as defined above, functional groups present in a
compound of formula II, with the exception of the groups participating in
the reaction, being, if necessary, in protected form, or a salt of such a
compound is reacted with a compound of formula III
##STR3##
wherein R.sub.2 is as defined above, functional groups present in a
compound of formula III, with the exception of the guanidino group
participating in the reaction, being, if necessary, in protected form, or
with a salt of such a compound, and any protecting groups present are
removed, or
b) for the preparation of a compound of formula I wherein R.sub.1 is
pyridyl, pyrazinyl thiazolyl, pyrimidinyl, pyridazinyl or imidazolyl, each
of which is substituted by a radical of the formula --N(R.sub.7)--R.sub.8,
and R.sub.2 has any one of the above-mentioned meanings, a compound of
formula I wherein R.sub.1 is pyridyl, pyrazinyl, thiazolyl, pyrimidinyl,
pyridazinyl or imidazolyl, each of which is substituted by a leaving
group, is reacted with an amine of formula
HN(R.sub.7) (IV),
wherein the substituents are as defined above, functional groups present
in a compound of formula IV, with the exception of the amino group
participating in the reaction, being, if necessary, in protected form, and
any protecting groups present are removed, or
c) for the preparation of a compound of formula I wherein R.sub.1 is any
one of the above-mentioned cyclic radicals substituted by carbamoyl or by
a radical of the formula --C(.dbd.O)--OR.sub.3, wherein R.sub.3 is
hydrogen, and R.sub.2 has any one of the above-mentioned meanings, a
compound of formula I wherein R.sub.1 is any one of the above-mentioned
cyclic radicals substituted by cyano is hydrolysed, or
d) for the preparation of a compound of fomula I nwherein R.sub.1 is a
pyridyl radical substituted by cyano or by --OR.sub.9, wherein R.sub.9 is
hydrogen or lower alkyl, and R.sub.2 has any one of the above-mentioned
meanings, in an N-oxido-pyridyl compound of formula VIII
##STR4##
wherein R.sub.21 is N-oxido-pyridyl bonded to a ring carbon atom and
R.sub.2 has any one of the above-mentioned meanings, the N-oxido group is
converted into a leaving group and the resulting leaving group is removed
from the molecule by nucleophilic substitution in the ortho-position with
respect to the pyridyl nitrogen using a nucleophile that introduces
hydroxy, cyano or unsubstituted or halogen-substituted lower alkoxy, or
e) for the preparation of a compound of formula I wherein R.sub.1 is a
pyridyl radical substituted by chlorine and R.sub.2 has any one of the
above-mentioned meanings, an N-oxido-pyridyl compound of formula VIII
##STR5##
wherein R.sub.21 is N-oxido-pyridyl bonded to a ring carbon atom and
R.sub.2 has any one of the above-mentioned meanings, is reacted with a
reagent that introduces chlorine in the ortho-position with respect to the
N-oxido group, and, if desired, a compound of formula I obtainable in
accordance with any one of Process a-e is converted into its salt, or an
obtainable salt of a compound of formula I is converted into the free
compound.
The manner in which the above-mentioned process variants are carried out is
explained in detail hereinafter.
General
The end products of formula I may comprise substituents that can also be
used as protecting groups in starting materials for the preparation of
other end products of formula I. Within the scope of this text, therefore,
unless the context indicates otherwise, only a readily removable group
that is not a constituent of the particular end product of formula I
desired is referred to as a "protecting group".
Protecting groups and the manner in which they are introduced and removed
are described, for example, in "Protective Groups in Organic Chemistry",
Plenum Press, London, New York 1973, and in "Methoden der organischen
Chemie", Houben-Weyl, 4th edition, Vol. 15/1, Georg-Thieme-Verlag,
Stuttgart 1974 and in Theodora W. Greene, "Protective Groups in Organic
Synthesis", John Wiley & Sons, New York 1981. A characteristic of
protecting groups is that they can be readily removed, that is to say,
without undesired secondary reactions taking place, for example by
solvolysis, reduction, photolysis or also under physiological conditions.
Hydroxy-protecting groups are, for example, acyl radicals, such as
unsubstituted or substituted, for example halogen-substituted, lower
alkanoyl, such as 2,2-dichloroacetyl, or acyl radicals of carbonic acid
semiesters, especially tert-butoxycarbonyl, unsubstituted or substituted
benzyloxycarbonyl, for example 4-nitrobenzyloxycarbonyl, or
diphenylmethoxycarbonyl, or 2-halo-lower alkoxycarbonyl, such as
2,2,2-trichloroethoxycarbonyl, and also trityl or formyl, or organic silyl
or stannyl radicals, and also readily removable etherifying groups, such
as left-lower alkyl, for example tert-butyl, 2-oxa- or 2-thia-aliphatic or
-cycloaliphatic hydrocarbon radicals, especially 1-lower alkoxy-lower
alkyl or 1-lower alkylthio-lower alkyl, for example methoxymethyl,
1-methoxy-ethyl, 1-ethoxyethyl, methylthiomethyl, 1-methylthioethyl or
1-ethylthioethyl, or 2-oxa- or 2-thia-cycloalkyl having 5 or 6 ring atoms,
for example tetrahydrofuryl or 2-tetrahydropyranyl or corresponding this
analogues, and also unsubstituted or substituted 1-phenyl-lower alkyl,
such as unsubstituted or substituted benzyl or diphenylmethyl, suitable
substituents of the phenyl radicals being, for example, halogen, such as
chlorine, lower alkoxy, such as methoxy, and/or nitro.
A protected amino group may, for example, be in the form of a readily
cleavable acylamino, arylmethylamino, etherified mercaptoamino,
2-acyl-lower alk-1-en-yl-amino, silylamino or stannylamino group or in the
form of an azido group.
In a corresponding acylamino group, acyl is, for example, the acyl radical
of an organic carboxylic acid having, for example, up to 18 carbon atoms,
especially of an alkanecarboxylic acid that is unsubstituted or
substituted, for example, by halogen or by aryl, or of a benzoic acid that
is unsubstituted or substituted, for example, by halogen, lower alkoxy or
by nitro, or of a carbonic acid semiester. Such acyl groups are, for
example, lower alkanoyl, such as formyl, acetyl or propionyl, halo-lower
alkanoyl, such as 2-halo-acetyl, especially 2-chloro-, 2-bromo-, 2-iodo-,
2,2,2-trifluoro- or 2,2,2-trichloro-acetyl, benzoyl that is unsubstituted
or substituted, for example, by halogen, lower alkoxy or by nitro, for
example benzoyl, 4-chlorobenzoyl, 4-methoxybenzoyl or 4-nitrohenzoyl, or
lower alkoxycarbonyl that is branched in the 1-position of the lower alkyl
radical or suitably substituted in the 1- or 2-position, especially
tert-lower alkoxycarbonyl, for example tert-butoxycaxbonyl,
arylmethoxycarbonyl having one or two aryl radicals that are preferably
phenyl that is unsubstituted or mono- or poly-substituted, for example, by
lower alkyl, especially left-lower alkyl, such as tert-butyl, lower
alkoxy, such as methoxy, hydroxy, halogen, for example chlorine, and/or by
nitro, such as unsubstituted or substituted benzyloxycarbonyl, for example
4-nitrobenzyloxycarbonyl, or substituted diphenylmethoxycarbonyl, for
example benzhydryloxycarbonyl or di(4-methoxyphenyl)methoxycarbonyl,
aroylmethoxycarbonyl wherein the aroyl group is preferably benzoyl that is
unsubstituted or substituted, for example, by halogen, such as bromine,
for example phenacyloxycarbonyl, 2-halo-lower alkoxycarbonyl, for example
2,2,2-trichloroethoxycarbonyl, 2-bromoethoxycarbonyl or
2-iodoethoxycarbonyl, or 2-(trisubstituted silyl)ethoxycarbonyl wherein
the substituents are each independently of the others an aliphatic,
araliphatic, cycloaliphatic or aromatic hydrocarbon radical that is
unsubstituted or substituted, for example, by lower alkyl, lower alkoxy,
aryl, halogen or by nitro, and contains up to 15 carbon atoms, such as
corresponding unsubstituted or substituted lower alkyl, phenyl-lower
alkyl, cycloalkyl or phenyl, for example 2-tri-lower
alkylsilylethoxycarbonyl, such as 2-trimethylsilylethoxycarbonyl or
2-(di-n-butyl-methyl-silyl)-ethoxycarbonyl, or
2-triarylsilylethoxycarbonyl, such as 2-triphenylsilylethoxycarbonyl.
Other acyl radicals suitable as amino-protecting groups are also
corresponding radicals of organic phosphoric, phosphonic or phosphinic
acids, such as di-lower alkylphosphoryl, for example dimethylphosphoryl,
diethylphosphoryl, di-n-propylphosphoryl or diisopropylphosphoryl,
dicycloalkylphosphoryl, for example dicyclohexylphosphoryl, unsubstituted
or substituted diphenylphosphoryl, for example diphenylphosphoryl,
unsubstituted or substituted, for example nitro-substituted,
di(phenyl-lower alkyl)phosphoryl, for example dibenzylphosphoryl or
di(4-nitrobenzyl)phosphoryl, unsubstituted or substituted
phenyloxyphenylphosphonyl, for example phenyloxyphenylphosphonyl, di-lower
alkylphosphinyl, for example diethylphosphinyl, or unsubstituted or
substituted diphenylphosphinyl, for example diphenylphosphinyl.
In an arylmethylamino group that is a mono-, di- or, especially,
tri-arylmethylamino group, the aryl radicals are especially unsubstituted
or substituted phenyl radicals. Such groups are, for example, benzyl-,
diphenylmethyl- and, especially, trityl-amino.
An etherified mercapto group in an amino group protected by such a radical
is especially arylthio or aryl-lower alkylthio wherein aryl is especially
phenyl that is unsubstituted or substituted, for example, by lower alkyl,
such as methyl or tert-butyl, lower alkoxy, such as methoxy, halogen, such
as chlorine, and/or by nitro. A corresponding amino-protecting group is,
for example, 4-nitrophenylthio.
In a 2-acyl-lower alk-1-en-1-yl radical that can be used as an
amino-protecting group, acyl is, for example, the corresponding radical of
a lower alkanecarboxylic acid, of a benzoic acid that is unsubstituted or
substituted, for example, by lower alkyl, such as methyl or tert-butyl,
lower alkoxy, such as methoxy, halogen, such as chlorine, and/or by nitro,
or especially of a carbonic acid semiester, such as a carbonic acid lower
alkyl semiester. Corresponding protecting groups are especially 1-lower
alkanoyl-prop-1-en-2-yl, for example 1-acetyl-prop-1-en-2-yl, or 1-lower
alkoxycarbonyl-prop-1-en-2-yl, for example 1 -ethoxycarbonyl-prop- 1
-en-2-yl.
Preferred amino-protecting groups are acyl radicals of carbonic acid
semiesters, especially tert-butoxycarbonyl, benzyloxycarbonyl that is
unsubstituted or substituted, for example, as indicated, for example
4-nitro-benzyloxycarbonyl, or diphenylmethoxycarbonyl, or 2-halo-lower
alkoxycarbonyl, such as 2,2,2-trichloroethoxycarbonyl, and also trityl or
formyl. The removal of the protecting groups that are not constituents of
the desired end product of formula I is effected in a manner known per se,
for example by solvolysis, especially hydrolysis, alcoholysis or
acidolysis, or by means of reduction, especially hydrogenolysis or
chemical reduction, as appropriate stepwise or simultaneously.
A protected amino group is freed in a manner known per se and, depending on
the nature of the protecting groups, in various manners, preferably by
solvolysis or reduction. 2-halo-lower alkoxycarbonylamino (where
appropriate after conversion of a 2-bromo-lower alkoxycarbonylamino group
into a 2-iodo-lower alkoxycarbonylamino group), aroylmethoxycarbonylamino
or 4-nitrobenzyloxycarbonylamino can be cleaved, for example, by treatment
with a suitable chemical reducing agent, such as zinc in the presence of a
suitable carboxylic acid, such as aqueous acetic acid.
Aroylmethoxycarbonylamino can also be cleaved by treatment with a
nucleophilic, preferably salt-forming reagent, such as sodium
thiophenolate, and 4-nitro-benzyloxycarbonylamino also by treatment with
an alkali metal dithionite, for example sodium dithionite. Unsubstituted
or substituted diphenylmethoxycarbonylamino, left-lower
alkoxycarbonylamino or 2-tri-substituted silylethoxycarbonylamino can be
cleaved by treatment with a suitable acid, for example formic acid or
trifluoroacetic acid, unsubstituted or substituted benzyloxycarbonylamino,
for example, by hydrogenolysis, that is to say by treatment with hydrogen
in the presence of a suitable hydrogenation catalyst, such as a palladium
catalyst, unsubstituted or substituted triarylmethylamino or formylamino,
for example, by treatment with an acid, such as a mineral acid, for
example hydrochloric acid, or an organic acid, for example formic, acetic
or trifluoroacetic acid, where appropriate in the presence of water, and
an amino group protected by an organic silyl group can be freed, for
example, by hydrolysis or alcoholysis. An amino group protected by
2-haloacetyl, for example 2-chloroacetyl, can be freed by treatment with
thiourea in the presence of a base, or with a thiolate salt, such as an
alkali metal thiolate, of the thiourea, and subsequent solvolysis, such as
alcoholysis or hydrolysis, of the resulting condensation product. An amino
group protected by 2-substituted silylethoxycarbonyl can also be converted
into the free amino group by treatment with a hydrofluoric acid salt
yielding fluoride anions.
A hydroxy group protected by a suitable acyl group, an organic silyl group
or by unsubstituted or substituted 1-phenyl-lower alkyl is freed
analogously to a correspondingly protected amino group. Hydroxy protected
by unsubstituted or substituted 1-phenyl-lower alkyl, for example benzyl,
is freed preferably by catalytic hydrogenation, for example in the
presence of a palladium-on-carbon catalyst. A hydroxy group protected by
2,2-dichloroacetyl is freed, for example, by basic hydrolysis, and a
hydroxy group etherified by tert-lower alkyl or by a 2-oxa- or
2-thia-aliphatic or -cycloaliphatic hydrocarbon radical is freed by
acidolysis, for example by treatment with a mineral acid or a strong
carboxylic acid, for example trifluoroacetic acid. Hydroxy etherified by
an organic silyl radical, for example trimethylsilyl, can also be freed by
a hydrofluoric acid salt yielding fluoride anions, for example
tetrabutylammonium fluoride.
Process a
Preferably, R17 and R.sub.18 are each methyl.
Free functional groups in a compound of formula II, which are
advantageously protected by readily removable protecting groups, are
especially amino groups in the radical R.sub.l.
Free functional groups in a compound of formula III, which are
advantageously protected by readily removable protecting groups, are
especially amino groups, but also hydroxy and carboxy groups.
A salt of a compound of formula II or III is preferably an acid addition
salt, for example a nitrate or one of the acid addition salts mentioned
for the end products of formula I.
The reaction is carried out in a suitable solvent or dispersing agent, for
example a suitable alcohol, such as 2-methoxy-ethanol or a suitable lower
alkanol, for example isopropanol or isobutanol, at a temperature of from
room temperature (approximately 20.degree. C.) to 150.degree. C., for
example under reflux. Especially when the compound of formula II is used
in the form of a salt, that salt is converted into the free compound,
preferably in situ, by the addition of a suitable base, such as an alkali
metal hydroxide, for example sodium hydroxide.
The starting material of formula II is obtained by reacting a compound of
formula V
##STR6##
wherein R.sub.1 is as defined above, with a compound of formula VI
##STR7##
wherein R.sub.19 and R.sub.20 are each lower alkyl and the other
substituents are as defined above, analogously to the procedure described
in the European Patent Application having the publication number 233 461.
Typical representatives of a compound of formula VI are
N,N-dimethylformamide dimethylacetal and N,N-dimethylformamide
diethylacetal. The reaction is effected while heating the reactants of
formulae V and VI, for example for 1-24 hours, in the absence or, if
necessary, in the presence of a solvent, at a temperature of approximately
from 50.degree. C. to 150.degree. C.
Alternatively, the starting material of formula II can also be obtained by
reacting a compound of formula V with formic acid ethyl ester of the
formula H--C(.dbd.O)--O--CH.sub.2 --CH.sub.3 and reacting the resulting
product with an amine of the formula H--N(R.sub.17)--R.sub.18 wherein the
substituents are as defined above.
The starting material of formula III is obtained in the form of an acid
addition salt by reacting an aniline derivative of formula VII
##STR8##
wherein R.sub.2 is as defined above, with cyanamide (NC--NH.sub.2). The
reaction is effected in a suitable solvent or dispersing agent, for
example a suitable alcohol, for example a suitable lower alkanol, such as
ethanol, for example
.alpha.) in the presence of equimolar amounts of the salt-forming acid, for
example nitric acid,
or
.beta.) in the presence of a clear, for example 60%, excess of a mineral
acid, such as hydrochloric acid, an ammonium salt of the desired
salt-forming acid, for example ammonium nitrate, being added when the
reaction is complete,
at a temperature of from room temperature to 150.degree. C., for example
under reflux.
Process b
A leaving group is reactive esterified hydroxy, for example hydroxy
esterified by a strong inorganic or organic acid, such as by a mineral
acid, for example a hydrohalic acid, such as hydrochloric, hydrobromic or
hydriodic acid, also sulfuric acid or a sulfuryl halide, for example
sulfuryl fluoride, or by a strong organic sulfonic acid, such as a lower
alkanesulfonic acid that is unsubstituted or substituted, for example, by
halogen, such as fluorine, or an aromatic sulfonic acid, for example a
benzenesulfonic acid that is unsubstituted or substituted by lower alkyl,
such as methyl, halogen, such as bromine, and/or by nitro, for example a
methanesulfonic, trifluoromethanesulfonic or p-toluenesulfonic acid. A
preferred leaving group is halogen, such as, especially, chlorine.
The reaction is preferably carried out in the presence of an excess of the
amine of formula IV, which can, where appropriate, also be used as
solvent, and, if necessary, in the presence of an inert solvent, such as
dimethyl sulfoxide, at a temperature of from room temperature to
+150.degree. C., for example at 100.degree. C.
Process c
The hydrolysis of cyano to carbamoyl can be carried out in the presence of
a suitable weak base, such as an alkali metal carbonate, for example
sodium carbonate. In order to prevent the hydrolysis from continuing
partially to carboxy, it is recommendable to carry out the hydrolysis with
hydrogen peroxide in the presence of a suitable olefin, such as preferably
a lower alkene, for example 1-hexene, in the presence of an alkali metal
carbonate, for example sodium carbonate, in a suitable solvent, such as an
alcohol, such as preferably ethanol, at more temperature.
The hydrolysis of cyano to carboxy is carried out in a suitable solvent,
such as an alcohol, such as ethanol, for example in the presence of a
suitable base, such as aqueous sodium hydroxide solution, at temperatures
of from room temperature to +150.degree. C., for example at 60.degree. C.
Process d
The conversion of the N-oxide group into a leaving group is effected, for
example, by reaction with a suitable reactive carboxylic or sulfonic acid
derivative, for example with a suitable lower alkanoic acid chloride,
lower alkanoic acid anhydride, such as acetic anhydride,
N,N-dimethyl-carbamoyl chloride, toluenesulfonyl chloride, methanesulfonyl
chloride or trifluoromethanesulfonyl chloride. A nucleophile that
introduces cyano is, for example, a suitable silyl cyanide, such as
tri-lower alkyl-silyl cyanide, for example tri-methylsilyl cyanide. A
nucleophile that introduces lower alkoxy or halogen-substituted lower
alkoxy is, for example, a corresponding lower alkanol, or a suitable metal
salt, such as, for example, an alkali metal salt, thereof, that is to say,
a corresponding lower alkanolate. Hydroxy can be introduced, for example,
by reacting a compound of formula VIII with a suitable acid anhydride and
hydrolysing the resulting intermediate.
Process d is carried out in a suitable solvent, such as acetonitrile, at
temperatures of approximately from 0.degree. C. to 150.degree. C.,
preferably approximately from room temperature to 100.degree. C.
The starting material of formula VIII is obtained, for example, by
oxidising a corresponding pyridyl compound analogous to formula VIII,
wherein R.sub.21 is pyridyl bonded to a ring carbon atom, with a suitable
oxidising agent, such as hydrogen peroxide or a suitable peracid, for
example a suitable perbenzoic acid, such as especially m-chloro-perbenzoic
acid, in an inert solvent, such as methylene chloride, at room
temperature.
Alternatively, the starting material of formula VIII can be obtained, for
example, by first oxidising acetyl-pyridine, such as 4-acetyl-pyridine,
with m-chloro-perbenzoic acid in a suitable solvent, such as methylene
chloride, for example under reflux, to acetyl-pyridine N-oxide, such as
4-acetyl-pyridine N-oxide, then converting the resulting acetyl-pyridine
N-oxide, such as 4-acetyl-pyridine N-oxide, with dimethylformamide
diethylacetal, which, for example, simultaneously serves as solvent, for
example at approximately 110.degree. C., into
3-dimethylamino-1-(N-oxido-pyridyl)-2-propen-1-one, such as
3-dimethylamino-1-(N-oxido-4-pyridyl)-2-propen-1-one, and then reacting
the latter with an R.sub.2 -phenyl-guanidine wherein R.sub.2 is as defined
above, or preferably with a suitable salt, for example a nitrate, thereof
in a suitable solvent, such as isopropanol, and in the presence of a
suitable base, such as sodium hydroxide, for example under reflux, to form
a compound of formula VIII.
In another method of preparing the starting material of formula VIII, the
above-mentioned acetyl-pyridine N-oxide, such as 4-acetyl-pyridine
N-oxide, is converted with phosphorus oxychloride in a suitable inert
solvent, such as toluene, for example at approximately 100.degree. C.,
first into acetyl-2-chloro-pyridine, for example
4-acetyl-2-chloro-pyridine. The resulting acetyl-2-chloro-pyridine, for
example 4-acetyl-2-chloro-pyridine, is then converted with
dimethylformamide diethylacetal, which, for example, simultaneously serves
as solvent, for example at approximately 110.degree. C., into
3-dimethylamino-1-(2-chloro-pyridyl)-2-propen-1-one, such as
3-dimethylamino-1-(2-chloro-4-pyridyl)-2-propen-1-one, which is then
reacted with a suitable salt, for example a nitrate, of an R.sub.2
-phenyl-guanidine, wherein R.sub.2 is as defined above, in a suitable
solvent, such as isopropanol, and in the presence of a suitable base, such
as sodium hydroxide, for example under reflux, to form a compound of
formula VIII.
Process e
A reagent that introduces chlorine in the ortho-position with respect to
the N-oxido group is, for example, phosphorus pentachloride,
trifluoromethylsulfonyl chloride/HCl gas or preferably phosphorus
oxychloride. By reacting a compound of formula VIII (for preparation see
above under Process d) with such a reagent, such as especially phosphorus
oxychloride, a compound of formula I is obtained wherein R.sub.1 is a
chlorine-substituted pyridyl radical that no longer contains an N-oxido
group. The reaction with phosphorus oxychloride can be carried out, for
example, in the absence of a solvent at approximately 100.degree. C.
Alternatively, it is possible to use phosphorus oxychloride together with
a suitable amine, such as diisopropylamine, in a suitable solvent, for
example a chlorinated hydrocarbon, such as chloroform, at approximately
room temperature. Another possibility is to use phosphorus oxychloride in
a suitable solvent, such as chloroform, toluene or xylene, at elevated
temperature, for example under reflux.
Acid addition salts of compounds of formula I are obtained in customary
manner, for example by treatment with an acid or a suitable anion exchange
reagent.
Acid addition salts can be converted in customary manner into the free
compounds, for example by treatment with a suitable basic agent.
Mixtures of isomers can be separated into the individual isomers in a
manner known per se, for example by fractional crystallisation,
chromatography, etc.
The processes described above, including the processes for removing
protecting groups and the additional process measures are, unless
otherwise indicated, carried out in a manner known per se for example in
the presence or absence of preferably inert solvents or diluents, if
necessary in the presence of condensation agents or catalysts, at reduced
or elevated temperature, for example in a temperature range of from
approximately -20.degree. C. to approximately 150.degree. C., especially
from approximately 0.degree. C. to approximately +70.degree. C.,
preferably from approximately +10.degree. C. to approximately +50.degree.
C., principally at room temperature, in a suitable vessel and, if
necessary, in an inert gas atmosphere, for example a nitrogen atmosphere.
Taking into account all the substituents in the molecule, if necessary, for
example if readily hydrolysable radicals are present, especially mild
reaction conditions are to be used, such as short reaction times, the use
of mild acidic or basic agents in low concentration, stoichiometric
ratios, and the selection of suitable catalysts, solvents, temperature
conditions and/or pressure conditions.
The invention relates also to those forms of the process in which a
compound obtainable as intermediate at any stage of the process is used as
starting material and the remaining process steps are carried out or the
process is discontinued at any stage or a starting material is formed
under the reaction conditions or is used in the form of a reactive
derivative or salt. The starting materials used are preferably those
which, according to the process, result in the compounds described above
as being especially valuable.
The present invention relates also to novel starting materials and/or
intermediates and to processes for the preparation thereof. The starting
materials used and the reaction conditions chosen are preferably such that
the compounds described in this Application as being especially preferred
are obtained.
The invention relates also to a method of treating warm-blooded animals
suffering from a tumour disease, which method comprises administering to
warm-blooded animals requiring such treatment an amount that is effective
in inhibiting tumours of a compound of formula I or of a pharmaceutically
acceptable salt thereof. The invention relates also to the use of a
compound of formula I or of a pharmaceutically acceptable salt thereof in
the inhibition of protein kinase C in warm-blooded animals or in the
preparation of pharmaceutical compositions for use in the therapeutic
treatment of the human or animal body. Depending on the species, age,
individual condition, mode of administration and the particular clinical
picture, effective doses, for example daily doses of approximately 1-1000
mg, especially 50-500 mg, are administered to a warm-blooded animal of
approximately 70 kg body weight.
The invention relates also to pharmaceutical compositions comprising an
effective amount, especially an mount effective in the prophylaxis or
treatment of one of the above-mentioned disorders, of the active
ingredient together with pharmaceutically acceptable carriers that are
suitable for topical, enteral, for example oral or rectal, or parenteral
administration and that may be inorganic or organic, solid or liquid.
There are used for oral administration especially tablets or gelatin
capsules that comprise the active ingredient together with diluents, for
example lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or
glycerol, and/or lubricants, for example silica, talc, stearic acid or
salts thereof, such as magnesium or calcium stearate, and/or polyethylene
glycol.
Tablets may also comprise binders, for example magnesium aluminium
silicate, starches, such as corn, wheat or rice starch, gelatin,
methylcellulose, sodium carboxymethylcellulose and/or
polyvinylpyrrolidone, and, if desired, disintegrators, for example
starches, agar, alginic acid or a salt thereof, such as sodium alginate,
and/or effervescent mixtures, or adsorbents, dyes, flavourings and
sweeteners. It is also possible to use the pharmacologically active
compounds of the present invention in the form of parenterally
administrable compositions or in the form of infusion solutions. Such
solutions are preferably isotonic aqueous solutions or suspensions which,
for example in the case of lyophilised compositions that comprise the
active ingredient alone or together with a carrier, for example mannitol,
can be made up prior to use. The pharmaceutical compositions may be
sterilized and/or may comprise excipients, for example preservatives,
stabilisers, wetting agents and/or emulsifiers, solubilisers, salts for
regulating the osmotic pressure and/or buffers. The present pharmaceutical
compositions, which may, if desired, comprise other pharmacologically
active substances, such as antibiotics, are prepared in a manner known per
se, for example by means of conventional mixing, granulating,
confectioning, dissolving or lyophilising processes, and comprise
approximately from 1% to 100%, especially from approximately 1% to
approximately 20%, active ingredient(s).
The following Examples illustrate the invention without limiting it in any
way. The R.sub.f values are determined on silica gel thin-layer plates
(Merck, Darmstadt, Germany). The ratio of the eluants in the eluant
mixtures used is indicated in parts by volume (v/v) and temperatures are
indicated in degrees Celsius.
Abbreviations
HV: high vacuum
RT: room temperature
EXAMPLE 1
31 mg (0.78 mmol) of sodium hydroxide are added to a suspension of 150 mg
(0.7 mmol) of 3-dimethylamino-1-(2-chloro-4-pyridyl)-2-propen-1-one and
165 mg (0.71 mmol) of 3-chloro-phenyl-guanidine nitrate in 1.5 ml of
2-propanol. After stirring under reflux for 18 hours, the reaction mixture
is cooled and filtered and the material retained on the filter is washed
thoroughly with water. After drying (60.degree., HV),
N-(3-chloro-phenyl)-4-(2-chloro-4-pyridyl)-2-pyrimidineamine is obtained;
m.p. 196.degree.-198.degree., R.sub.f =0.67 (methylene
chloride:methanol=95:5), FAB-MS: 317 (M.sup.+ +1).
The starting material is obtained in the following manner:
Stage 1.1
4.12 ml (39.12 mmol) of 3-chloro-aniline are placed in 25 ml of ethanol,
and 3.3 g (78.4 mmol) of cyanamide are added. 5.3 ml (62.7 mmol) of
concentrated hydrochloric acid are added dropwise to the brown solution.
The reaction solution is then stirred for 20 hours at 78.degree.. After
concentration under reduced pressure, the residue is dissolved in 25 ml of
water, and 6.3 g (78.4 mmol) of ammonium nitrate are added. The
precipitated substance is isolated by filtration, washed with water and
dried at 60.degree. under HV. 3-chloro-phenyl-guanidine nitrate is
obtained; .sup.1 H-NMR (dimethyl sulfoxide): 7.2-7.8 (7H,m), 9.9
(1H,br,s).
Stage 1.2
24.61 g (177.62 mmol) of 2-chloro-4-cyano-pyridine are placed in 1.25
liters of diethyl ether under nitrogen, and 120 ml (22% in
tetrahydrofuran, 353 mmol) of methylmagnesium chloride are added. The red
suspension is stirred for 40 hours at RT, poured onto 1.25 liters of
ice/water and 250 ml of 6N hydrochloric acid and stirred for 14 hours at
RT. Extraction with diethyl ether and methylene chloride, drying with
MgSO.sub.4 and concentration give 4-acetyl-2-chloro-pyridine; R.sub.f =0.5
(methylene chloride:methanol=9:1).
Stage 1.3
16.2 g (104.2 mmol) of 4-acetyl-2-chloro-pyridine are stirred for 1 hour at
110.degree. with 116 ml of dimethylformamide diethylacetal. After cooling
to 0.degree., filtering and drying at 60.degree. under HV,
3-dimethylamino-1-(2-chloro-4-pyridyl)-2-propen-1-one is obtained; .sup.1
H-NMR (dimethyl sulfoxide): 2.98 (3H,s), 3.2 (3H,s), 5.9 (1H,d), 7.8
(3H,m), 8.5 (1H,d).
EXAMPLE 2
Analogously to Example 1 there is obtained from 150 mg (0.7 mmol) of
3-dimethylamino-1-(3-chloro4-pyridyl)-2-propen-1-one and 190 mg (0.71
mmol) of 3-tri-fluoromethyl-phenyl-guanidine nitrate
N-(3-trifluoromethyl-phenyl)-4-(2-chloro-4-pyridyl)-2-pyrimidineamine;
m.p. 168.degree.-171.degree., R.sub.f 0.67 (methylene
chloride:methanol=95:5).
The starting material is obtained in the following manner:
Stage 2.1
Analogously to Stage 1.1 there is obtained from 16.1 g (0.1 mol) of
3-trifluoromethyl-aniline and 6.3 g (0. 15 mol) of cyanamide
3-trifluoromethyl-phenyl-guanidine nitrate; .sup.1 H-NMR (DMSO): 7.6
(7H,m), 9.9 (1H,br,s).
EXAMPLE 3
0.8 g (2.41 mmol) of
N-(3-trifluoromethyl-phenyl)-4-(N-oxido4-pyridyl)-2-pyrimidineamine is
suspended in 40 ml of acetonitrile. 0.834 ml (6.65 mmol) of
tri-methylsilyl cyanide and 0.611 ml (6.665 mmol) of dimethylcarbamoyl
chloride are added and the reaction mixture is stirred for 12 hours at
60.degree.. After concentration under reduced pressure, crystallisation is
effected from tetrahydrofuran/diethyl ether.
N-(3-trifluoromethyl-phenyl)-4-(2-cyano-4-pyridyl)-2-pyrimidineamine is
obtained; m.p. 164.degree.-166.degree., R.sub.f =0.40 (n-hexane:ethyl
acetate=1:1).
The starting material is obtained in the following manner:
Stage 3.1
15.1 g (0.0567 mmol) of 3-trifluoromethyl-phenyl-guanidine nitrate and 2.84
g (70.9 mmol) of sodium hydroxide are added to a suspension of 10 g (56.7
mmol) of 3-dimethylamino-1-(4-pyridyl)-2-propen-1-one ›described in EP-A-0
233 461! in 300 ml of isopropanol The reaction mixture is boiled under
reflux for 24 hours. After cooling, the product is isolated by filtration,
washed with water and dried at 60.degree. under HV.
N-(3-trifluoromethyl-phenyl)-4-pyridyl-2-pyrimidineamine is obtained; m.p.
197.degree.-198.degree., R.sub.f =0.58 (ethyl acetate).
Stage 3.2
10.57 g (33.4 mmol) of
N-(3-trifluoromethyl-phenyl)-4-pyridyl-2-pyrimidineamine are suspended in
200 ml of methylene chloride, and 10.49 g (33.42 mmol, 55% strength) of
m-chloroperbenzoic acid are added. After 2 hours, 200 ml of water are
added. The reaction product is isolated by filtration, washed with sodium
carbonate solution and water and, after drying,
N-(3-trifluoromethyl-phenyl)-4-(N-oxido-4-pyridyl)-2-pyrimidineamine is
obtained. More product is obtained by chromatography (methylene
chloride:methanol=9: 1) of the concentrated mother liquor; R.sub.f =0.16
(methylene chloride:methanol=9:1).
EXAMPLE 4
100 mg (0.293 mmol) of
N-(3-trifluoromethyl-phenyl)-4-(2-cyano4-pyridyl)-2-pyrimidineamine are
stirred in 15 ml of ethanol and 15 ml of 2N sodium hydroxide solution for
3 hours at 60.degree.. After acidifying with 4N hydrochloric acid,
N-(3-trifluoromethyl-phenyl)-4-(2-carboxy-4-pyridyl)-2-pyrimidineamine is
obtained; m.p. 241.degree.-245.degree., FAB-MS: 361 (M.sup.+ +H).
EXAMPLE 5
500 mg (1.67 mmol) of
N-(3-chloro-phenyl)-4-(N-oxido-4-pyridyl)-2-pyrimidineamine are suspended
in 5 ml of acetonitrile, and 0.42 ml (4.5 mmol) of dimethylcarbamoyl
chloride and 0.56 ml (4.5 mmol) of trimethylsilyl cyanide are added. After
stirring for 14 hours at 60.degree., the reaction mixture is cooled to RT
and the reaction product is isolated by filtration and washed with diethyl
ether. Recrystallisation from tetrahydrofuran gives
N-(3-chloro-phenyl)-4-(2-cyano-4-pyridyl)-2-pyrimidineamine in the form of
yellow crystals; m.p. 221.degree.-222.degree., R.sub.f =0.6 (hexane:ethyl
acetate=1:1).
The starting material is obtained in the following manner:
Stage 5.1
2.8 g (12 mmol) of 3-chloro-phenyl-guanidine nitrate and 0.5 g (12 mmol) of
sodium hydroxide are added to a suspension of 2.0 g (11.7 mmol) of
3-dimethylamino-1-(4-pyridyl)-2-propen-1-one ›described in EP-A-0 233 461!
in 100 ml of isobutanol and the reaction mixture is boiled under reflux
for 5 hours. After cooling, the reaction product is isolated by
filtration, washed with water and chromatographed (tetrahydrofuran). After
crystallisation (tetrahydrofuran/diethyl ether),
N-(3-chloro-phenyl)-4-pyridyl-2-pyrimidineamine is obtained; m.p.
167.degree.-168.degree., R.sub.f =0.38 (methylene chloride:methanol=9:1).
Stage 5.2
1.0 g (3.54 mmol) of N-(3-chloro-phenyl)-4-(pyridyl)-2-pyrimidineamine are
suspended in 50 ml of methylene chloride, and 1.1 g of m-chloroperbenzoic
acid (50% strength) are added. After stirring for 18 hours at RT, the
reaction mixture is filtered, the residue is dissolved in ethyl
acetate/tetrahydrofuran (1:1) and extracted with 1N sodium hydroxide
solution and water. The dried organic phase is concentrated and the
residue is crystallised from diethyl ether/tetrahydrofuran to give
N-(3-chloro-phenyl)-4-(N-oxido-4-pyridyl)-2-pyrimidineamine; m.p.
268.degree.-270.degree., R.sub.f =0.6 (methylene chloride:methanol=9:1).
EXAMPLE 6
50 mg (0.16 mmol) of
N-(3-chloro-phenyl)-4-(2-cyano-4-pyridyl)-2-pyrimidineamine are stirred in
5 ml of ethanol and 5 ml of 2N sodium hydroxide solution for 2 hours at
60.degree.. After cooling to RT, the product is isolated by filtration and
washed with ethanol/water (9:1 ) and dried at 50.degree. under HV. The
sodium salt of
N-(3-chloro-phenyl)-4-(2-carboxy-4-pyridyl)-2-pyrimidineamine is obtained;
m.p. >250.degree., R.sub.f =<0.1 (methylene chloride:methanol=9:1).
EXAMPLE 7
50 mg (0.16 mmol) of
N-(3-chloro-phenyl)-4-(2-cyano-4-pyridyl)-2-pyrimidineamine are suspended
in 2 ml of methanol. 0.58 ml of hydrogen peroxide (30% strength), 0.16 ml
of 1-hexene and 11 mg of sodium carbonate are added and the reaction
mixture is stirred for 14 hours at RT. The product is isolated by
filtration, washed (methanol:water=1:1) and dried at 50.degree. under HV.
N-(3-chloro-phenyl)-4-(2-carbamoyl-4-pyridyl)-2-pyrimidineamine is
obtained in the form of a yellow powder; m.p. 245.degree.-247.degree.,
R.sub.f =0.23 (n-hexane:ethyl acetate=1:1).
EXAMPLE 8
100 mg (0.293 mmol) of
N-(3-trifluoromethyl-phenyl)-4-(2-cyano-4-pyridyl)-2-pyrimidineamine are
suspended in 4 ml of methanol. 1.1 ml of hydrogen peroxide (30%), 0.32 ml
of 1-hexene and 22 mg of sodium carbonate are added and the reaction
mixture is stirred for 16 hours at RT. The product is isolated by
filtration and washed (methanol/water) to give
N-(3-trifluoromethyl-phenyl)-4-(2-carbamoyl-4-pyridyl)-2-pyrimidineamine;
m.p. 240.degree.-242.degree., FAB-MS: 360 (M.sup.+ +H).
EXAMPLE 9
In a manner analogous to that described above and by simple conversion
reactions, known per se, of the products, the following compounds are
prepared:
a) N-(3-chloro-phenyl)-4-(2-n-propylamino4-pyridyl)-2-pyrimidineamine,
b) N-(3-chloro-phenyl)-4-(2-amino-4-pyridyl)-2-pyrimidineamine,
c) N-(3-chloro-phenyl)-4-(2-hydroxy-4-pyridyl)-2-pyrimidineamine and
d) N-(3-chloro-phenyl)-4-(2-methoxy-4-pyridyl)-2-pyrimidineamine.
EXAMPLE 10
300 mg (0.95 mmol) of
N-(3-chloro-phenyl)-4-(2-chloro-4-pyridyl)-2-pyrimidineamine (see Example
1), 5.3 ml of 1,3-propanediol and 3.0 ml of dimethylformamide are stirred
for 43 hours at 105.degree.. After concentration and repeated
chromatography (methylene chloride:methanol=98:2)
N-(3-chloro-phenyl)-4-(2-dimethylamino-4-pyridyl)-2-pyrimidineamine is
obtained; m.p. 176.degree.-178.degree., FAB-MS: 326 (M.sup.+ +H).
EXAMPLE 11
14.5 g (53.7 mmol) of 3-ethoxycarbonyl-phenyl-guanidine nitrate, 11.3 g
(53.7 mmol) of 3-dimethylamino-1-(2-chloro4-pyridyl)-2-propen-1-one and
2.4 g (60 mmol) of sodium hydroxide are stirred in 150 ml of isobutanol
for 14 hours at 110.degree.. After cooling, washing twice with 100 ml of
ethanol each time and crystallising (tetra-hydrofuran/ethanol),
N-›3-ethoxycarbonyl-phenyl!-4-(2-chloro4-pyridyl)-2-pyrimidineamine is
obtained; m.p. 149.degree.-150.degree., FAB-MS: 355 (M.sup.+ +H).
EXAMPLE 12
N-›3-isopropoxycarbonyl-phenyl!-4-(2-chloro-4-pyridyl)-2-pyrimidineamine is
isolated as a secondary product of Example 11; m.p.
130.degree.-131.degree., FAB-MS: 383 (M.sup.+ +H).
EXAMPLE 13
9.4 g (26.5 mmol) of
N-›3-ethoxycarbonyl-phenyl!-4-(2-chloro-4-pyridyl)-2-pyrimidineamine (see
Example 11) and 50 ml of 2N sodium hydroxide solution are boiled under
reflux in 300 ml of ethanol for 1 hour. After cooling to RT, the reaction
mixture is acidified (4N hydrochloric acid) and the reaction product is
isolated by filtration. After drying at 50.degree. under HV, lemon-yellow
crystals of N-›3-carboxy-phenyl!-4-(2-chloro-4-pyridyl)-2-pyrimidineamine
are obtained; m.p. 267.degree.-268.degree., FAB-MS: 327 (M.sup.+ +H).
EXAMPLE 14
Analogously to Example 1 there is obtained from 100 mg (0.32 mmol) of
N-›3-chloro-phenyl!-4-(2-chloro-4-pyridyl)-2-pyrimidineamine and 3 ml
(29.9 mmol) of n-1-butylamine N-›3-chloro-phenyl!-4-›2-(n-1
-butylamino)-4-pyridyl!-2-pyrimidineamine; m.p. 151.degree.-158.degree.,
FAB-MS: 354 (M.sup.+ +H).
EXAMPLE 15
Tablets each comprising 20 mg of active ingredient, for example one of the
compounds of formula I described in Examples 1-14, are prepared with the
following composition in customary manner:
______________________________________
Composition:
______________________________________
active ingredient 20 mg
wheat starch 60 mg
lactose 50 mg
colloidal silica 5 mg
talc 9 mg
magnesium stearate 1 mg
145 mg
______________________________________
Preparation
The active ingredient is mixed with a portion of the wheat starch, with the
lactose and with the colloidal silica, and the mixture is forced through a
sieve. A further portion of the wheat starch is made into a paste with 5
times the amount of water on a water bath, and the powder mixture is
kneaded with the paste until a slightly plastic mass has been formed.
The plastic mass is pressed through a sieve of approximately 3 mm mesh size
and dried, and the resulting dry granules are forced through a sieve
again. The remainder of the wheat starch, the talc and the magnesium
stearate are admixed and the mixture is compressed to form tablets each
weighing 145 mg and having a breaking notch.
EXAMPLE 16
Capsules each comprising 10 mg of active ingredient, for example one of the
compounds of formula I described in Examples 1-14, are prepared in
customary manner as follows:
______________________________________
Composition:
______________________________________
active ingredient 2500 mg
talc 200 mg
colloidal silica 50 mg
______________________________________
Preparation
The active ingredient is intimately mixed with the talc and the colloidal
silica, and the mixture is forced through a sieve 0.5 mm mesh size and
introduced in 11-mg portions into hard gelatin capsules of suitable size.
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